Objective: The DIVA study is aimed at setting up a standardized genotypic tropism-testing on proviral-DNA for the routine clinical diagnostic-laboratory Methods: Twelve local centres and 5 reference centres (previously cross-validated) were identified. For inter-center validation-procedure, 60 peripheral-blood mononuclear cells (PBMCs) aliquots from 45 HAART-treated patients were randomly chosen for population V3 sequencing on proviral-DNA at local HIV centre and at reference-laboratory Viral tropism was predicted by Geno2Pheno algorithm (False Positive Rate [FPR] = 20%) as proposed by the European-Guidelines. Quantification of total HIV-1 DNA was based on a method described by Viard (2004). Results: Quantification of HIV-1 DNA was available for 35/45 (77.8%) samples, and gave a median value of 598 (IQR:252-1,203) copies/10(6) PBMCs. A total of 56/60 (93.3%) samples were successfully amplified by both the reference and the local virological centers. The overall concordance of tropism prediction between local and reference centers was 54/56 (96.4%). Results of tropism prediction by local centers were: 33/54 (61.1%) R5 and 21/54 (38.9%) X4/DM. Conclusion: There was high concordance in the genotypic tropism prediction based on proviral DNA among different virological centers throughout Italy. Our results are in line with other European studies, and support the use of genotypic tropism testing on proviral DNA in patients with suppressed plasma HIV-1 RNA candidate to CCR5-antagonist treatment.

Performance of genotypic tropism testing on proviral DNA in clinical practice: results from the DIVA Study Group.

BORDERI, MARCO;BON, ISABELLA;RE, MARIA CARLA;
2012

Abstract

Objective: The DIVA study is aimed at setting up a standardized genotypic tropism-testing on proviral-DNA for the routine clinical diagnostic-laboratory Methods: Twelve local centres and 5 reference centres (previously cross-validated) were identified. For inter-center validation-procedure, 60 peripheral-blood mononuclear cells (PBMCs) aliquots from 45 HAART-treated patients were randomly chosen for population V3 sequencing on proviral-DNA at local HIV centre and at reference-laboratory Viral tropism was predicted by Geno2Pheno algorithm (False Positive Rate [FPR] = 20%) as proposed by the European-Guidelines. Quantification of total HIV-1 DNA was based on a method described by Viard (2004). Results: Quantification of HIV-1 DNA was available for 35/45 (77.8%) samples, and gave a median value of 598 (IQR:252-1,203) copies/10(6) PBMCs. A total of 56/60 (93.3%) samples were successfully amplified by both the reference and the local virological centers. The overall concordance of tropism prediction between local and reference centers was 54/56 (96.4%). Results of tropism prediction by local centers were: 33/54 (61.1%) R5 and 21/54 (38.9%) X4/DM. Conclusion: There was high concordance in the genotypic tropism prediction based on proviral DNA among different virological centers throughout Italy. Our results are in line with other European studies, and support the use of genotypic tropism testing on proviral DNA in patients with suppressed plasma HIV-1 RNA candidate to CCR5-antagonist treatment.
Svicher V; Alteri C; Montano M; D'Arrigo R; Andreoni M; Angarano G; Antinori A; Antonelli G; Allice T; Bagnarelli P; Baldanti F; Bertoli A; Borderi M; Boeri E; Bon I; Bruzzone B; Callegaro AP; Capobianchi MR; Carosi G; Cauda R; Ceccherini-Silberstein F; Clementi M; Chirianni A; Colafigli M; D'Arminio Monforte A; De Luca A; Di Biagio A; Di Nicuolo G; Di Perri G; Di Pietro M; Di Santo F; Fabeni L; Fadda G; Galli M; Gennari W; Ghisetti V; Giacometti A; Gori C; Gori A; Gulminetti R; Leoncini F; Maffongelli G; Maggiolo F; Manca G; Gargiulo F; Martinelli C; Maserati R; Mazzotta F; Meini G; Micheli V; Monno L; Mussini C; Narciso P; Nozza S; Paolucci S; Pal G; Parisi S; Parruti G; Pignataro AR; Pollicita M; Quirino T; Re MC; Rizzardini G; Santangelo R; Scaggiante R; Sterrantino G; Turriziani O; Vatteroni ML; Vecchi L; Viscoli C; Vullo V; Zazzi M; Lazzarini A; Perno CF.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11585/114016
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